Idiotypic vaccine for treatment of human B-cell lymphoma. Construction of IgG variable regions from single malignant B cells.
Immunoglobulin idiotypes (Id) of malignant B cells represent highly specific markers which can be used for vaccination. PCR-amplification of immunoglobulin genes enables the rapid production of large amounts of Id vaccines. However, the separate amplification and subsequent recombination of heavy and light chains can lead to a loss of the relevant Id. To preserve the original chain pairs, we used single malignant B cells derived from an immunocytoma patient. Cytoplasm was extracted and the mRNA transcribed into cDNA. The VH and VL genes were then amplified by PCR and cloned into a vector for expression in E. coli. Id production was checked using an anti-Id mouse monoclonal Ab raised against the patient's tumor-specific IgG. One out of 3 constructs expressed the relevant Id. Analysis of the first 31 light chain residues revealed an identical sequence for the malignant B cells' IgG and the recombinant Id construct. Exchange of either the heavy or light chain with an unrelated chain resulted in loss of the Id. An unrelated sequence derived from the c-myc protein is coupled to the Id vaccine. The lymphoma patient was shown to have Abs to the c-myc sequence. This sequence therefore, increases the Id+ Ab's antigenicity. CD spectroscopy showed an alpha-helical structure for the c-myc epitope. In conclusion, a B-cell lymphoma autovaccine was produced containing immunogenic sequences that do not alter the steric conformation of the tumor-specific Id.
Terness
P
P
Institute of Immunology, University of Heidelberg, Germany.
Welschof
M
M
Moldenhauer
G
G
Jung
M
M
Moroder
L
L
Kirchhoff
F
F
Kipriyanov
S
S
Little
M
M
Opelz
G
G
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000096:biosynthesis / Q000627:therapeutic use; D000951:Antigens, Neoplasm / Q000032:analysis; D019496:Cancer Vaccines / Q000276:immunology / Q000627:therapeutic use*; D000939:Epitopes / Q000097:blood; D016259:Genes, myc / Q000276:immunology; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000032:analysis; D007130:Immunoglobulin Idiotypes; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis; D016393:Lymphoma, B-Cell / Q000097:blood / Q000276:immunology / Q000628:therapy*; D051379:Mice; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D011487:Protein Conformation; D017421:Sequence Analysis; D014614:Vaccines, Synthetic / Q000096:biosynthesis
D000888:Antibodies, Anti-Idiotypic; D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D000939:Epitopes; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007130:Immunoglobulin Idiotypes; D007135:Immunoglobulin Variable Region; D010455:Peptides; D014614:Vaccines, Synthetic